Tonon SA, Ferreras JA, Liotta DJ, Bos PD, Galuppo J, Zinovich JB

Language: English
References: 24
Page: 117-120
PDF size: 55.62 Kb.

ABSTRACT

Epidemiological studies have identified human papilloma virus (HPV) infection as the main risk factor associated with development of cervical cancer. It has been found that 80 to 100% of advanced neoplasms and cervical invasive carcinomas contain only a few HPV types. Type 16 is the most prevalent of the invasive carcinomas (30-60%). It has therefore been suggested that follow-up and treatment of infected patients must be directed against the specific infective virus. Our objective was to develop a rapid method of identifying HPV 16 in cervical smears. To do this, a type-specific PCR amplification stage has been designed for the region corresponding to the HPV 16 E6 oncogene, followed by product confirmation by restriction enzyme digestion analysis. This method has been applied to fifteen fresh cytological cervical smear samples, obtained from patients infected with HPV 16, which were previously diagnosed by HSIL. The CaSki cell line genome carrying HPV 16 was used as the positive control and the HeLa cell line genome, normal human lymphocyte genome and two cytological samples infected with HPV 18 and 6, respectively were used as negative controls. A specific 140 bp amplification product was obtained from the HPV 16 E6 oncogene in all samples analyzed and in the positive control. Further enzyme restriction with Rsa I produced the expected cutting pattern of two fragments consisting of 90 and 50 bp. Negative controls and the samples infected with HPV 18 and 6 did not produce a specific amplification product. This method could be used as a rapid method of identifying HPV 16 in fresh cervical samples.

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